Three dimensional growth of animal cell by new immobilization technique and its application for artificial liver

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In order to establish artificial liver, long term passage of liver cells has been tried. Using liver cells isolated by collegenase method, effects of several vitamins, and amino acids for cell maintenance were studied. Among them, proline which is main component of collagen, and activated ascorbate which is a cofactor of collagen biosynthesis, facilitated albumin biosynthesis at least for one week. On the other hand, super oxide dismutase which decompose toxic super oxide anion also facilitate albumin secretion.We also studied the suitable materials for immobilization of liver cells and developed a macroporous fiber which bound to heparin or collagen as cell attachment ligands. Vero cell attached very well to both of matrix and cell density was more than 10^7/ml support. This result indicated that the support may be suitable for artificial liver construction.For the construction of artificial blood vessel, we isolated human endothelial cells. By the infection of a temperature sensitive mutant of SV40 virus, life span of cells was elongated for more than 200days at 33ﾟC. These cells died at 37ﾟC since the temperature sensitive tumor protein was inactivated at this temperature. Since these cells expressed endothelial cell specific function, they are useful for the analyses of blood vessel related disorder.